[Construction of recombinant fowlpox virus coexpressing HA from subtype H5 of avian influenza virus and chicken interleukin-18].
OBJECTIVE
We developed recombinant fowlpox viruses (rFPV) coexpressing chicken IL-18 and H5 AIV HA.


METHODS
Recombinant expression plasmid pSYHA/IL-18 was constructed by cloning chicken IL-18 into transfer plasmid containing HA gene and transfected by lipofectamine on the chicken embryo fibroblasts cell (CEF) pre-infected with S-FPV-017. By selecting blue plaques on the CEF overlaid with agar containing X-gal, recombinants fowlpox virus rFPV-HA-IL-18 were obtained, and identified by PCR.


RESULTS
The recombinant fowlpox viruses contained chicken IL-18 and HA gene and had stable genetic properties. The expression of HA was detected in the recombinant virus-infected CEF by indirect immunofluorescence using antibody against AIV. The expression of chicken IL-18 was detected by MTT in the recombinant virus-infected CEF fluid. The chickens vaccinated with recombinant fowlpox virus rFPV-HA-IL-18 and rFPV-HA had detectable hemagglutination inhibition (HI) antibody at 7 days post-vaccination, and HI antibody titers rose to peak at 14 days post-vaccination. No HI antibody was detected in the control or fowlpox virus immunized chickens before or after immunization. The chickens vaccinated with rFPV-HA-IL-18 had higher HI antibody titers than the chickens vaccinated with rFPV-HA.


CONCLUSIONS
Development of recombinant fowlpox virus (rFPV-HA-IL-18) had strong biological activity.
Chen
Hongying
H
College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China. chhy927@163.com
Huang
Qingyun
Q
Cui
Bao'an
B
Li
Xinsheng
X
Guan
Qian
Q
D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D002645:Chickens; D004274:DNA, Recombinant / Q000235:genetics* / Q000276:immunology; D019084:Fluorescent Antibody Technique, Indirect; D005587:Fowlpox virus / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics; D006384:Hemagglutination / Q000276:immunology; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics* / Q000276:immunology; D009980:Influenza A virus / Q000737:chemistry / Q000145:classification / Q000235:genetics; D020382:Interleukin-18 / Q000235:genetics*; D007700:Kinetics; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism; D014611:Vaccination
D000906:Antibodies; D004274:DNA, Recombinant; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D020382:Interleukin-18; C109963:hemagglutinin, avian influenza A virus